Cell Phenotype-Driven Bioinks for 3D Disease Models

last updated: 2021-12-16
ProjectECM_INK :: publications list
TitleCell Phenotype-Driven Bioinks for 3D Disease Models
Publication TypeComunications - Poster
Year of Publication2019
AuthorsReis D. P., Malta M. D., Gasperini L., Cerqueira M. T., Reis R. L., and Marques A. P.

Bioprinting, as a tissue engineering tool, enables the deposition of various cells and matrices at high resolution, accuracy and speed. These features represent a major advantage to recreate the different microenvironments/microfunctionalities found in each tissue which enlightens the potential of these biofabrication method for the development of physiologically reliable 3D in vitro tissue models. In 3D printing, it is necessary to use materials with superior shear-thinning and recovery properties, such as Gellan Gum (GG), to develop printable bioinks. To render physiological resemblance to the material, the aim of this work was to combine GG with biological components produced by skin cells of different phenotypes. For that purpose, dystrophic epidermolysis bullosa (DEB)fibroblasts were selected as a tool, to develop a 3D model capable of mimicking the native diseased tissue. In the future, these models can be used to further study the disease and contribute to the development of new therapies. Healthy and dystrophic epidermolysis bullosa (DEB) diseased fibroblasts were cultured and their protein content was extracted, characterized and conjugated with GG, to produce disease-specific bioinks when combined with cells. Cells were cultured for 21 days in culture medium supplemented with 50 µg/ml ascorbic acid. An optimised protocol was followed to extract the maximum amount of the ECM produced. Extracts were combined with GG to form hydrogels. Preliminary results demonstrated the possibility to conjugate components from hDFbs ECM to GG, obtaining stable gels under in vitro conditions for up to 14 days. During this timeframe, proteins released from the gels were quantified. A gradual release of protein content occurred within the first 7 days, while the structural integrity of the gels was maintained. Regarding cell studies, 1x104 hDFBs were encapsulated within the gels, and their cytocompatibility was assessed by Live/Dead and Phalloidin/DAPI staining. Good cell viability was observed and cytoskeletal staining showed that the amount of ECM components determines cell adhesion and morphology. These results confirmed that it is possible to combine an otherwise inert polymer (GG) with ECM produced from cells of different pathological phenotypes, conferring biological cues to this hydrogel with potential to develop bioinks for 3D printing of different disease models. Acknowledgments: FCT Doctoral Program in Tissue Engineering, Regenerative Medicine and Stem Cells PD/BD/14301/2018 (DPR); SFRH/BD/137766/2018 (MDM); CEECIND/00695/2017 (MTC), EB house Austria, and Consolidator Grant Project “ECM_INK” (ERC-2016-COG-726061).

Conference NameTERM STEM 2019
Date Published2019-11-01
Conference LocationBraga
Keywords3D disease models, Bioinks
Peer reviewedno

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